THE DISRUPTION OF CAVEOLAE/CAVEOLIN-1 PRESENT IN TROPHOBLAST CELLS DID NOT AFFECT ENDOTHELIAL CELL MIGRATION

NORA MARTÍNEZ; JULIETA REPPETTI; ALEJANDRA RECA; ABRIL SEYAHIAN; NATALIA SZPILBARG; ALICIA E DAMIANO

Chillan

Congreso; II MEETING ON RESEARCH AND INNOVATION IN VASCULAR HEALTH; 2017

THE DISRUPTION OF CAVEOLAE/CAVEOLIN-1 PRESENT INTROPHOBLAST CELLS DID NOT AFFECT ENDOTHELIAL CELLMIGRATIONMartínez Nora1, Reppetti Julietta 1, Reca Alejandra 1, Seyahian Abril1, Szpilbarg Natalia 1Damiano Alicia E 1,21Laboratory of Biology of Reproduction, Institute of Physiology and Biophysics Bernardo Houssay (IFIBIO)- CONICET- School of Medicine, Universidad de Buenos Aires, Buenos Aires, Argentina.2Departament of Biological Sciences, School of Pharmacy and Biochemistry, Universidad de Buenos Aires, Buenos Aires, Argentina.Migration plays key roles in many physiological and pathological processes including development, angiogenesis, tissue regeneration and metastasis. Caveolin-1 (Cav-1) and caveolae constitute domains that compartmentalize intracellular signalling pathways. During placentation the human extravillous trophoblast (EVT) cells invade the endometrium and maternal vasculature within the uterus.Objective: To evaluate the role of caveolae/Cav-1 in the migration and invasion oftrophoblast cells and the interaction with the endothelial cells.Methods: Cav-1 expression was examined in trophoblast cells Swan-71 by immunocytochemistry and western blot. Cells were treated with 10 mM methyl-β-cyclodextrin (β-MCD) to disrupt caveolae. Conditioned medium (CM) was also collected. Migration was assessed by wound healing assay at 8, 18 and 24 h. To evaluate invasion, trophoblast cells were seeded in the top of transwell inserts of 8-μm pore precoated with Matrigel (0.3 mg/ml). After 24 h, cells located on the undersurface were fixed with methanol and stained with hematoxylin. To evaluate the interaction with the endothelial cells (EA.hy926), proliferation (evaluated by MTT assay) and migration wereperformed after the culture of the cells with the CM obtained from Swan-71 cells.Results: The migration of Swan-71 treated with β-MCD was decreased compared with control: 8 h (32.78 ± 1.14 vs 17.78 ± 0.5561), 18 h (66.74 ± 1.434 vs 51.79 ± 1.158), 24 h(84.45 ± 1.160 vs 64.95 ± 0.7165). In the Invasion assay in Swan-71 no differences were observed between control and β-MCD treated cells. No differences were also found in the proliferation and in the migration of endothelial cells treated with the CM from Swan-71.Conclusion: These results show that lipid raft disruption significantly attenuates migration (but no invasion) in Swan-71 cells suggesting a role for Caveolae/caveolin-1during the early stages of placental development. Nevertheless the lack of caveolae/caveolin-1 in the trophoblast cells did not affect the proliferation and the migration of endothelial cells.